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71.
Presence of extrachromosomal DNa in Rhodopseudomonas capsulata strain BH9 was shown by the appearance of a satellite band in a dye-buoyant density gradient. Radioactively labelled DNA was prepared from this satellite band and examined on a 5–20% sucrose gradient. Three radioactive peaks with sedimentation coefficients of 100 S, 94 S, and 58–64 S, respectively, were consistently observed. Analysis of these sedimentation coefficients suggested that there are two species of plasmid DNA with molecular sizes of 94×106 daltons (named pBH91) and 74×106 daltons (named pBH92). The 58–64 S peak is attributed to open circular molecules. DNAs from each peak of the sucrose gradient were examined by electronmicroscopy, and the results agree closely with those of the sucrose gradient analysis. Reassociation kinetics of the plasmid DNA was also followed. Addition of total DNA of strain BH9 increased the renaturation rate of the plasmid DNA. It was calculated from the magnitude of the increase that approximately 10% of the BH9 total DNA may hybridize with the plasmid sequences. DNA prepared from the gene transfer agent (GTA) produced by R. capsulata increases the renaturation rate of the plasmid to the same extent as total DNA isolated from the GTA producing strain, Y262. 相似文献
72.
The requirement of muscle phosphorylase for branched polysaccharide substrates was investigated by kinetic studies on semisynthetic branched saccharides. One series of saccharides was prepared from maltoheptose by oxidizing the reducing group to a carboxyl group and coupling this with an amino group of ethylenediamine. The resulting aminooligosaccharide was coupled with p-nitrophenyl esters of mono-, di-, tetra-, and polycarboxylic aicds to produce saccharides containing one, two, four, and approximately 52 maltodextrin chains per molecule. A similar series of saccharides was prepared from a heterogeneous maltodextrin of average chain length 11.7. Kinetic constants were determined for the reaction with phoshorylase a in the direction of chain elongation. Michaelis constants are equilibrium constants for dissociation of saccharide from the enzyme-AMP-glucose-1P-saccharide complex. The Michaelis constants, expressed in terms of the concentration of nonreducing end groups, are independent of maltodextrin chain length but decrease considerably as the number of chains per molecule increases. Maximum velocities do not differ greatly from that for glycogen. Among the synthetic saccharides, only the polymer behaves similarly to glycogen in exhiiting a decreasing reaction rate as the chains are elongated. The kinetic constants are quantitatively consistent with a model in which two chain termini from the same saccharide molecule bind to the phosphorylase molecule simultaniously, Differences in binding between saccharides having different numbers of equally accessible chains are caused solely by statistical factors in the equilibrium. Highly branched substrates bind better because of their greater multiplicity of two end-group pairs. 相似文献
73.
Jinchun Wu Ziye Zou Yang Liu Xuhao Liu Zhengrong Zhangding Mo Xu Jiazhi Hu 《Nucleic acids research》2022,50(19):11128
CRISPR/Cas9 has been adapted to disrupt endogenous genes in adoptive T-lymphocyte therapy to prevent graft-versus-host disease. However, genome editing also generates prevalent deleterious structural variations (SVs), including chromosomal translocations and large deletions, raising safety concerns about reinfused T cells. Here, we dynamically monitored the progression of SVs in a mouse model of T-cell receptor (TCR)-transgenic T-cell adoptive transfer, mimicking TCR T therapeutics. Remarkably, CRISPR/Cas9-induced SVs persist and undergo clonal expansion in vivo after three weeks or even two months, evidenced by high enrichment and low junctional diversity of identified SVs post infusion. Specifically, we detected 128 expanded translocations, with 20 615 as the highest number of amplicons. The identified SVs are stochastically selected among different individuals and show an inconspicuous locus preference. Similar to SVs, viral DNA integrations are routinely detected in edited T cells and also undergo clonal expansion. The persistent SVs and viral DNA integrations in the infused T cells may constantly threaten genome integrity, drawing immediate attention to the safety of CRISPR/Cas9-engineered T cells mediated immunotherapy. 相似文献
74.
Qiaoyu Chen Yanyan Hu Lijun Yang Benguo Zhu Feng Luo 《Current issues in molecular biology》2022,44(9):4070
Phosphorus treatment can reduce Cd accumulation and Cd toxicity in rice, but alterations in the internal regulatory network of rice during this process have rarely been reported. We have removed the effect of cadmium phosphate precipitation from the hydroponic system, treated a pair of different Cd-response rice varieties with different levels of phosphorus and cadmium and examined the changes in physiological indicators and regulatory networks. The results demonstrated that phosphorus treatment significantly reduced Cd accumulation in both types of rice, although the antioxidant systems within the two types of rice produced opposite responses. Overall, 3 mM phosphorus treatment to Cd-N decreased the expression of OsIAA17 and OsACO1 by 32% and 37%, respectively, while increasing the expression of OsNR2 by 83%; these three genes regulate the synthesis of auxin, ethylene, and nitric oxide in rice. IAA and NO levels in rice shoots increased by 24% and 96%, respectively, and these changes contribute to Cd detoxification. The cadmium transporter genes OsHMA2, OsIRT1, and OsABCC1 were significantly down-regulated in Cd-N roots after triple phosphorus treatment. These data suggest that phosphorus treatment can reduce Cd accumulation and enhance Cd resistance in rice by affecting the expression of signaling molecules. 相似文献
75.
Daptomycin is a cyclolipopeptide antibiotic produced by Streptomyces roseosporus. It is widely used to treat drug-resistant bacterial infections; however, daptomycin yield in wild strains is very low. To improve the daptomycin production by the strain BNCC 342432, a modified method of ribosome engineering with superposition of streptomycin resistance was adopted in this study. The highest-yield mutant strain SR-2620 was obtained by increasing streptomycin resistance of BNCC 342432, and achieved daptomycin production of 38.5 mg/l in shake-flask fermentation, 1.79-fold higher than the parent strain and its heredity stability was stable. The morphological characteristics of the two strains were significantly different, and the 440th base G of the rpsL gene in the mutant strain was deleted, which resulted in a frameshift mutation. Our results demonstrate that gradually increasing strain resistance to streptomycin was an effective breeding method to improve daptomycin yield in S. roseosporus. Open in a separate window 相似文献
76.
花绒寄甲(Dastarcus helophoroides)(鞘翅目:寄甲科Bothrideridae)是寄生栗山天牛(Massicus raddei)中老龄幼虫和蛹的重要天敌,但其寄主栗山天牛世代周期长(3年1代)、发育比较整齐,不利于寄生性天敌种群数量的稳定.为了解利用花绒寄甲防治栗山天牛后,其种群能否在栎树林间长期保持较高的种群数量,达到持续控制栗山天牛的防治效果,调查研究了花绒寄甲在栎树林间的转主寄主和种群保持机制.结果表明,在东北辽东栎树干和树枝上除了栗山天牛外,还有其他8种天牛危害:双簇天牛(Moechotypa diphysis)、四点象天牛(Mesosa myops)、中华薄翅锯天牛(Megopis sinica)、锯天牛(Prionus insularis)、双带粒翅天牛(Lamiomimus gottschei)、八字绿虎天牛(Chlorophorus tohokensis)、日本绿虎天牛(C.japonicus)和拟蜡天牛(Stenygrinumquadrinotatum).其中以栗山天牛、双簇天牛、四点象天牛和拟蜡天牛数量较多,而花绒寄甲在辽东栎树干上的垂直分布与栗山天牛、双簇天牛和四点象天牛的垂直分布重叠较多.室内研究表明,花绒寄甲对四点象天牛老熟幼虫的寄生率达到26.67%,对蛹的寄生率达到了43.33%;对双簇天牛老熟幼虫的寄生率达到20.00%,对蛹的寄生率为6.67%.对双簇天牛和四点象天牛在林间的生活史调查和研究发现,花绒寄甲可寄生的这两种天牛的中老龄幼虫和蛹,在花绒寄甲不适宜寄生的栗山天牛幼龄幼虫期大量存在,表明双簇天牛和四点象天牛是花绒寄甲在栎树林中的主要转主寄主.由于这些转主寄主的存在,花绒寄甲在不利于其寄生的栗山天牛卵期、幼龄幼虫期可转移寄生这些寄主,从而在栗山天牛危害的栎树林间保持了较高的种群数量,达到对栗山天牛长期而有效的持续控制效果. 相似文献
77.
78.
Ornithine decarboxylase-antizyme was induced in mammary gland of fasted lactating rats by administration of 1,3-diaminopropan-2-ol. Antizyme from mammary gland showed similar chemical and kinetic behavior to that previously reported by Canellakis and co-workers for antizyme from liver [J. S. Heller, W. F. Fong, and E. S. Canellakis (1976) Proc. Natl. Acad. Sci. USA 72, 1858-1862]; specifically the inhibitor was nondialyzable, heat labile, and ribonuclease insensitive, and the inhibition was time independent, proportional to the concentration of antizyme present, and noncompetitive with respect to the substrate, ornithine. However, ornithine decarboxylase-antizyme from mammary gland eluted from Sephadex G-75 with an apparent molecular mass of 55 kDa, compared with 27 kDa, for antizyme from liver under identical conditions. The elution pattern was unaffected by the presence of high salt concentrations, indicating that the larger size was not due to macromolecular complexes. The presence of antizyme-ornithine decarboxylase complex was detected in mammary gland of untreated lactating rats fasted for 6 or 24 h, thus indicating that antizyme plays a role in the regulation of ornithine decarboxylase in mammary gland under physiological conditions. 相似文献
79.
The distribution pattern of neuropeptide Y-like immunoreactivity (NPY-Li) in cat hypothalamus was studied using avidin-biotin modification of immunocytochemical method. This study showed cell bodies containing NPY-Li in the periventricular and the infundibular nuclei and also a moderate number of neurons with NPY-Li in the ventromedial nucleus, an observation not reported in earlier studies. Fibers with NPY-Li were noted throughout the hypothalamus, but most prominently within the periventricular regions. The location of NPY cells within the hypothalamus suggests the possibility of an interaction with dopaminergic and other proopiomelanocortinergic neurons. 相似文献
80.
Hu Q. Budinoff C. R. Liu G. X. Sommerfeld M. & Westerhoff P. 《Journal of phycology》2003,39(S1):24-24
Upper Klamath Lake (UKL) is the largest lake in Oregon (area 287 km2 , avg. depth 4.2 m). It is naturally eutrophic and regularly suffers nuisance summer blooms of cyanobacteria, principally Aphanizomenon flos-aquae (AFA). Sediment coring studies show that AFA was absent or minimal until about 1880 when a steady increase began, culminating in the blooms of recent decades. These studies show concomitant increases in sediment N (∼20%) and P (∼50%) along with shifts in the algal flora indicating increased eutrophication. These changes correlate with increased human impacts, such as deforestation, construction, roadbuilding etc., and especially the ditching, diking and draining of adjacent wetlands for conversion to agriculture. Agricultural nutrient runoff, especially P, has been often cited as the cause of the AFA blooms, and most attention has been focused on the dynamics of UKL during the summer bloom. We propose that a more significant factor may be the loss of early-season suppression of AFA because of the loss of the lake-associated wetlands, which originally constituted 42% of the lake area, and which have declined in area by 66.3% since the late 1800's. The melting of snow and ice in the spring would flush into the lake a surge of wetland plant decomposition products, most significantly organic acids and humic substances. We propose that formerly these wetland effluents caused a complex of effects on lake pH, solar UV transparency, photochemical interactions, nutrient availability, and Daphnia grazing dynamics, which would have combined to prevent the development of any AFA bloom. 相似文献